人工合成六倍体小麦与普通小麦杂交后代衍生群体的PPO基因分析

四倍体小麦与节节麦杂交培育的人工合成六倍体小麦已广泛应用于国内外小麦品种改良。以引自CIMMYT的Syn768、Syn769、Syn780和Syn786人工合成六倍体小麦分别与中国四川成都平原主栽普通小麦品种杂交、回交的BC2F2：6后代群体中选育的113份优良高代系和川麦38、川麦42、川麦43和川麦47育成品种为材料，采用SSR特异引物Xgwm312标记位点的PAGE凝胶电泳对其PPO基因型进行了研究。结果表明，Xgwm312位点的标记具有多态性，其PCR扩增产物可产生198bp、216bp、232bp和240bp四种等位基因变异片段。在所检测的117份后代衍生群体材料中，65份材料具有198bp片段的等位基因，占全部材料的55．56％；13份含216bp片段的等位基因，其频率为11．11％，35份材料具有232bp片段的等位基因，分布频率为29．91％；只有4份材料含有240bp片段的等位基因，仅占全部材料的3．42％。从每个人工合成六倍体小麦亲本材料所形成的后代衍生群体来看，基因等位变异片段分布频率各不相同。说明PPO基因在小麦杂交后代材料中基因型的表现存在着随机性，与亲本的基因型状况关系极大，并且在后代材料中出现了亲本都没有的240bp等位基因变异片段的材料。通过研究人工合成六倍体小麦与普通小麦杂交后代的PPO基因型，有助于提高分子标记育种效率，也有助于PPO基因型的多态性研究，并为人工合成六倍体小麦在我国小麦品种改良和分子标记育种中的应用提供依据和方法。     

人工合成小麦与普通小麦杂交后代衍生群体的Rht8基因分析

【目的】四倍体小麦与节节麦杂交培育的人工合成小麦已广泛应用于国内外小麦品种改良。通过研究人工合成小麦与普通小麦杂交后代的Rht8基因型,有助于提高分子标记育种效率,也有助于Rht8 基因型的多态性研究,并为人工合成小麦在中国小麦品种改良和分子标记育种中的应用提供依据和方法;【方法】以引自CIMMYT的人工合成小麦分别与中国四川成都平原主栽普通小麦品种杂交、回交的BC2F2:6后代群体中选育的113份优良高代系和川麦38、川麦42、川麦43和川麦47育成品种为材料,采用特异引物的PCR 扩增和改进的聚丙烯酰胺凝胶电泳对其Rht8基因型进行了研究;【结果】在以syn768、Syn769、Syn780和Syn786人工合成小麦为亲本的117份后代衍生群体检测材料中,Rht8基因型频率为77.78%。从每一个人工合成小麦形成的小的后代衍生群体看,Rht8基因型频率各不相同。以syn768为亲本的后代衍生群体,Rht8基因型频率最高,为96.70%;在以syn769为亲本而育成的优良高代系和川麦38、川麦42与川麦43育成品种中,Rht8基因型频率最低,为71.64%;以Syn780为亲本的后代衍生群体中,Rht8基因型频率为73.68%,分离比率约为3:1;以Syn786为亲本育成的材料只有川麦47,该品种不含有Rht8该基因;【结论】不论父本或母本的Rht8的基因型状况如何,它们所产生的杂交后代材料Rht8基因的遗传是随机的。     

人工合成六倍体小麦后代衍生群体遗传多样性检测

通过四倍体二粒小麦和节节麦杂交而获得的人工合成六倍体小麦,含有丰富的普通小麦品种改良有益基因,作为拓宽普通栽培小麦性状和新品种改良的新的种质资源已广泛应用于普通小麦的遗传改良实践中.利用分布于小麦A、B、D基因组21条染色体、28个不同染色体臂上的37对微卫星引物,对人工合成六倍体小麦与四川成都平原普通栽培小麦主栽品种杂交、回交经多代选择而形成的117份人工合成六倍体小麦衍生后代高代群体系(其中川麦38、川麦42、川麦43和川麦47为审定品种)进行了DNA分子水平上的分析,共检测到256个等位变异,平均每个SSR标记位点检测到6.92个等位变异,变幅在1到14之间.A、B、D基因组中,D基因组表现出的多态性信息含量最低,为0.4276,B基因组次之,为0.5346,A基因组最高,达到0.6145(A＞B＞D).辛普森指数比较的结果也反映出相同的变化趋势,A基因组最高,为1.1874,B基因组次之,为1.0810,D基因组最小,为0.8046(A＞B＞D).综合多态性信息含量和辛普森指数的估值,表明这一批人工合成六倍体小麦衍生后代群体接受的遗传基因既来自人工合成六倍体小麦,又来自普通栽培小麦,显示杂合度类型丰富,具有较高的遗传差异.根据SSR位点获得的等位基因变异片断的分布情况进行UPGMA聚类,发现A、B、D基因组基凶型间的遗传相似系数较低,A、B、D三个基因组所得平均遗传相似系数为0.4721,其中A基因组平均遗传相似系数为0.3797,B基因组平均遗传相似系数为0.4627,D基因组上平均遗传相似系数为0.5815,反映人工合成六倍体小麦后代衍生材料的遗传多样性处于较高水平.研究结果证明利用人工合成六倍体小麦所具有的普通小麦野生近缘种中的基因库改良现代小麦,丰富其遗传基础,减少其生物和非生物胁迫的脆弱性,是一条行之有效的途径.     

人工六倍体小麦后代衍生群体遗传多样性研究(英文)

通过四倍体二粒小麦和节节麦杂交而获得的人工合成六倍体小麦,含有丰富的普通小麦品种改良有益基因,作为拓宽普通栽培小麦性状和新品种改良的新的种质资源已广泛应用于普通小麦的遗传改良实践中.利用分布于小麦A、B、D基因组21条染色体、28个不同染色体臂上的37对微卫星引物,对人工合成六倍体小麦与四川成都平原普通栽培小麦主栽品种杂交、回交经多代选择而形成的117份人工合成六倍体小麦衍生后代高代群体系(其中川麦38、川麦42、川麦43和川麦47为审定品种)进行了DNA分子水平上的分析,共检测到256个等位基因变异,平均每个SSR标记位点检测到6.92个等位变异基因.每个SSR位点等位基因变异数在1～14个,变异幅度较大,表明SSR分子标记在人工合成六倍体小麦中表现出高水平的遗传变异.A,B,D基因组中,D基因组表现出的多态性信息含量最低,为0.427 6,B基因组次之,为0.534 6,A基因组最高,达到 0.614 5(A>B>D).辛普森指数比较的结果也反映出相同的变化趋势,A基因组最高,为1.187 4,B基因组次之,为1.081,D基因组最小,为0.804 6(A>B>D).综合多态性信息含量和辛普森指数的估值,表明这一批人工合成六倍体小麦后代衍生高代群体接受的遗传基因既来自人工合成六倍体小麦,又来自普通栽培小麦,显示杂合度类型丰富,具有较高的遗传差异.根据获得的等位基因变异片断的分布情况进行UPGMA聚类,发现A,B,D基因组基因型间的遗传相似系数较低, A,B,D三基因组37个SSR标记位点所得平均遗传相似系数为0.472 1,A基因组平均遗传相似系数为0.379 7,B基因组平均遗传相似系数为0.462 7,D基因组上平均遗传相似系数为0.581 5,反映出人工合成六倍体小麦后代衍生群体材料的遗传多样性处于较高水平.本研究结果证明利用人工合成六倍体小麦所具有的普通小麦野生近缘种中的基因库改良现代小麦,丰富其遗传基础,减少其生物和非生物胁迫的脆弱性,是一条行之有效的途径.     

人工合成六倍体小麦与其亲本间淀粉酶活性的比较

研究小麦多倍化初期幼苗中α-淀粉酶和β-淀粉酶的活性变化,以模拟普通小麦的人工合成异源六倍体小麦为材料,采用3,5-二硝基水杨酸（DNS）法分别测定发芽96.5h的人工合成六倍体小麦（第五代）及其亲本中这两种淀粉酶的活性。结果表明,人工合成六倍体小麦中淀粉酶的活性显著低于其亲本中该酶的活性。     

小麦未减数配子基因的连锁标记及染色体区段检测

六倍体普通小麦(Triticumaestivum L., AABBDD, 2n=42)由四倍体小麦(T. turgidum, AABB, 2n=28)与节节麦(Aegilops tauschii Cosson, DD, 2n=14)天然杂交,然后通过染色体自动加倍形成。加倍过程主要受四倍体小麦未减数配子基因控制,且不同四倍体小麦存在不同的遗传效应。本研究利用位于3B染色体上未减数配子基因QTug.sau-3B的连锁SSR标记Xgpw1146和高通量DArTseq分子标记,筛选出可能转入四倍体小麦未减数配子基因的人工合成小麦改良后代。在105份改良材料中检测出17份具有四倍体小麦的Xgpw1146等位位点,表明四倍体小麦的未减数配子基因可能转入了这17份材料。利用DArTseq高通量标记技术分析人工合成小麦SHW-L1的88份改良后代,发现含四倍体小麦Xgpw1146等位位点的材料均具有来自SHW-L1、且可能包含Xgpw1146的一个染色体区段,表明未减数配子基因临近区域以一个区段传递到改良后代。这些人工合成小麦改良材料在加倍单倍体育种中有重要的应用潜力。     

山西冬小麦品种Waxy蛋白分析及分子标记研究

利用聚丙烯酰胺凝胶电泳(SDS-PAGE)法分析了山西100份小麦品种(系)Waxy蛋白的缺失类型,筛选出缺失Wx-B1蛋白亚基的材料8份。利用2个STS标记和1个SSR标记对不同Waxy蛋白缺失类型进行鉴定,验证其在分子标记辅助育种中的有效性,结果表明:Wx-7A位点的STS引物在野生型中扩增出1条450 bp的特异带,而在缺失Wx-A1蛋白亚基的突变材料中扩增出1条327 bp的特异带;Wx-4A位点的STS引物在野生型中扩增出1条440 bp的特异带,缺失Wx-B1蛋白亚基的突变材料中没有该扩增片段;Wx-7A,Wx-7D位点的SSR引物在野生型中分别扩增出1条265 bp和1条204 bp的特异带,在缺失Wx-A1和Wx-D1蛋白亚基的突变材料中没有扩增出该特异带。这3个标记可以用于分子标记辅助育种。     

人工合成六倍体小麦在黄淮麦区育种中的利用性评价

为了解人工合成六倍体小麦（synthetic hexaploid wheat,SHW）在黄淮麦区育种改良中的应用价值,利用从国际玉米小麦改良中心引进的5份SHW与普通小麦驻麦305进行杂交、回交,对亲本及BC₂群体的主要农艺性状、产量相关指标和籽粒品质进行调查分析。结果表明,与普通小麦相比,SHW的株高、小穗数、穗粒数、收获系数和千粒重等是其不利性状,但具有较多的分蘖。SHW的纤维含量、面筋含量、蛋白含量和硬度都比驻麦305高（PAAAAA0.05）。BC₂的收获系数较SHW显著提升,其产量虽然比SHW有了较大幅度提升,但仍显著低于普通小麦。BC₂的蛋白质和面筋含量相对于普通小麦亲本显著提升,其中仅2018-2019年的BC₂-SHW1和BC₂-SHW2群体未达显著水平。这表明5份SHW可作为重要的品质改良资源应用于小麦育种中。     

我国部分小麦地方品种Waxy基因多样性研究

利用2对STS引物、1对SSR引物和1对基因特异引物分析了1739份中国小麦地方品种Waxy基因的变异情况。检测出Wx-Al基因缺失突变材料3份，Wx-Bl基因缺失突变材料25份，Wx-Dl基因缺失突变材料3份。利用SDS-PAGE方法对以上Waxy基因突变材料在蛋白质水平上进行鉴定，验证了Waxy分子标记在种质资源研究中的有效性。向育种家推荐一批具有不同Waxy基因缺失的地方种质，并提供了全部品种的缺失类型。     

四倍体小麦与六倍体小麦杂种的染色体遗传特性

四倍体栽培小麦(Triticum turgidum L., AABB)和普通小麦(Triticum aestivum L., AABBDD)是两种目前主要的小麦栽培种。通过远缘杂交转移利用四倍体小麦(或六倍体小麦)基因是六倍体小麦(或四倍体小麦)遗传改良的重要方法。然而,两者杂种F1为基因组组成不平衡的五倍体,其中A和B基因组染色体均为两套,而D基因组染色体仅一套。亲本间的遗传差异,包括核基因组和细胞质基因组,可能影响五倍体杂种的染色体传递效率。本研究以多个不同遗传背景的四倍体小麦和六倍体小麦为亲本,配置正反交五倍体杂种F1,采用多色荧光原位杂交技术分析自交F2代植株的染色体组成规律。结果表明,杂交亲本的遗传背景对杂种F1自交结实率影响显著;不论是以四倍体小麦还是六倍体小麦做母本, AB基因组染色体在F1自交过程中相对稳定, F2后代的数目均接近28条(27.9 vs. 28.0);以四倍体小麦为母本F2平均保留的D基因组染色体数显著多于以六倍体小麦为母本的后代(7.0 vs. 2.9)。因此,以四倍体小麦为最终目标后代时,应优先以六倍体小麦为母本进行杂交组合的配置;以六倍体小麦为最终目标后代时,应优先以四倍体小麦为母本开始最初的杂交组合配置。     

Allelic variation and composition of HMW-GS in advanced lines derived from d-genome synthetic hexaploid / bread wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.)

The objective of this study was to identify allelic variations at Glu-1 loci of wheat (Triticum aestivum L.) advanced lines derived from hybridization of bread wheat and synthetic hexaploid wheats (2n = 6x = 42; AABBDD). Locally adapted wheat genotypes were crossed with synthetic hexaploid wheats. From the 134 different cross combinations made, 202 F₈ advanced lines were selected and their HMW-GS composition was studied using SDS-PAGE. In total, 24 allelic variants and 68 HMW-GS combinations were observed at Glu-A1, Glu-B1, and Glu-D1 loci. In bread wheat, the Glu-D1 locus is usually characterized by subunits 1Dx2+1Dy12 and 1Dx5+1Dy10 with the latter having a stronger effect on bread-making quality. The subunit 1Dx5+1Dy10 was predominantly observed in these advanced lines. The inferior subunit 1Dx2+1Dy12, predominant in adapted wheat germplasm showed a comparative low frequency in the derived advanced breeding lines. Its successful replacement is due to the other better allelic variants at the Glu-D1 locus inherited in these synthetic hexaploid wheats from Aegilops tauschii (2n = 2x = 14; DD).     

川麦42遗传背景中人工合成小麦导入位点的SSR标记检测

硬粒小麦和节节麦是六倍体普通小麦的二级基因源,六倍体人工合成小麦遗传变异丰富、蕴藏着丰富的抗性基因,可供现代小麦改良利用。利用人工合成小麦基因资源与四川小麦杂交、回交,已育成了高产、优质、抗病小麦新品种“川麦42”。本文利用217对微卫星（SSR）引物检测“川麦42”遗传背景中人工合成小麦的导入位点,发现24个位点来源于人工合成小麦,占所用引物数的11.06%,远小于理论值25%。川麦42遗传背景中人工合成小麦导入位点在A、B和D染色体组分布频率不均衡,D组>B组>A组;人工合成小麦导入位点在川麦42各染色体间差异也很大,在1B、2B和5A染色体上分布较集中、片段较长,而在1A等11条染色体上则无导入位点;表明人工合成小麦的遗传位点并不按孟德尔遗传规律传至后代,人工选择压力导致遗传位点很大的偏分离行为。     

Polymorphism of waxy proteins in Iberian hexaploid wheats

A collection of 130 cultivars of bread wheat, 332 landraces of bread wheat and 144 spelt wheats was analysed for waxy proteins in the grain. The electrophoretic patterns showed very low polymorphism and most of the hexaploid wheats had the Wx-Ala, Wx-D1a and Wx-B1 alleles of ‘Chinese Spring’. Two alleles were detected at Wx-A1 (Wx-A1a, and Wx-A1b (null)), the latter was present in only 5.1% of the bread wheat landraces and 7.6% ofthe spelt wheats. No allelic variation was found at the Wx-D1 locus and all the hexaploid wheats had the Wx-D1a allele. Wx-B1 was the most polymorphic locus, with three alleles detected: Wx-B1a, Wx-B1b (null) and Wx-Blc coding for a Wx-B1 protein with a slightly different mobility from Wx-B1a. The null Wx-B1b allele was found in 10.8% of the bread wheat cultivars, 21.4% of the bread wheat landraces and 12.5% of the spelt wheats. Among the 604 hexaploid wheats analysed, only two bread wheat landraces (0.6%) and two spelt wheats (1.4%) had the null allele at both Wx-A1 and Wx-B1 loci.     

Assessment of genetic diversity in synthetic hexaploid wheats and their Triticum dicoccum and Aegilops tauschii parents using AFLPs and agronomic traits

Synthetic hexaploid wheats are of interest to wheat breeding programs, especially for introducing new genes that confer resistance to biotic and abiotic stresses. A group of 54 synthetic hexaploid wheats derived from crosses between emmer wheat(Triticum dicoccum, source of the A and B genomes) and goat grass (Aegilops tauschii, D genome donor) were investigated for genetic diversity. Using the AFLP technique, dendrograms revealed clear grouping according to geographical origin for the T. dicoccum parents but no clear groups for the Ae. tauschii parents. The geographical clustering of the T. dicoccum parents was also reflected in the dendrogram of their derived synthetic hexaploids. Diversity of the T. dicoccum parents and their derived synthetic hexaploids was further evaluated by measuring 18morphological and agronomic traits on the plants. Clustering based on morphological and agronomic data also reflected geographical origin. However, comparison of genetic distances obtained from AFLP and agronomic data showed no correlation between the two diversity measurements. Nevertheless, similarities among major clusters with the two systems could be identified. Based on percentage of polymorphic markers, the synthetic hexaploids had a considerably higher level of AFLP diversity (39%) than normally observed in cultivated hexaploid wheat (12–21%). This suggests that synthetic hexaploid wheats can be used to introduce new genetic diversity into the bread wheat gene pool. This revised version was published online in July 2006 with corrections to the Cover Date.     

Breeding for salinity resistance in mulberry (<Emphasis Type="Italic">Morus</Emphasis> spp.)

Late maturity α-amylase (LMA) is a genetic defect that is fairly widely spread in bread wheat (Triticum aestivum L.) germplasm, and recently detected in durum cultivars, which can result in unacceptably high α-amylase activity (low falling number) in ripe grain. LMA has also been observed at unexpectedly high frequency and severity in synthetic hexaploid wheats derived from the interspecific hybridisation of Triticum durum (AABB) and Aegilops tauschii (DD). Since synthetic hexaploids represent an important new source of resistances/tolerances to a range of biotic and abiotic stresses for wheat breeders, there is a pressing need to understand the mechanisms involved in LMA in synthetics and develop strategies for avoiding its adverse effects on grain quality. The objectives of this study were to firstly, compare the LMA phenotype of synthetics that varied for plant height, secondly, to characterise the LMA phenotype in groups of synthetics derived from the same durum parents and finally to determine whether LMA in primary synthetics is associated with the QTL previously reported in conventional bread wheat. More than 250 synthetic hexaploids, a range of durum cultivars and a doubled haploid population derived from Worrakatta (non-LMA) × AUS29663 (high LMA synthetic) were phenotyped and genotyped with markers reported to be linked to LMA in conventional bread wheat and markers diagnostic for the semi-dwarfing gene, Rht1. More than 85% of synthetics were prone to LMA, approximately 60% ranked as very high. Genetic control of LMA in synthetic hexaploids appeared to involve QTL located on 7B, and to a lesser extent 3B, similar to bread wheats. However, the LMA phenotype of many synthetic hexaploids appeared to be more extreme than could be explained by comparisons with bread wheat even taking into account the apparent absence of Rht1 in most genotypes. Other mechanisms, possibly triggered by the interaction between the AABB and DD genomes cannot be excluded. The presence of wild type rht1 in most synthetic hexaploids and their extreme height is difficult to reconcile with the semi-dwarf, Rht1, stature of many of the durums used in the interspecific hybridisation process. Mechanisms that could explain this observation remain unclear.     

Starch characterisation and variability in GBSS loci of synthetic hexaploid wheats and their durum and <Emphasis Type="Italic">Aegilops tauschii</Emphasis> parents

Greater variability in starch properties is found in lower ploidy wheats than in commercial hexaploid wheats. This paper reports on the starch properties and variability in granule bound starch synthase (GBSS) loci of 17 diploid (Aegilops tauschii) and 12 tetraploid (durums) potential progenitors of wheat, compared with 29 synthetic hexaploid wheats produced from such progenitors. Starch properties examined were granule size distribution, swelling power, amylose content, gelatinisation and amylose-lipid dissociation properties. A PCR screening method was able to detect the presence or absence of each of the three GBSS genes. It also detected polymorphisms in eight diploids and nine hexaploids, all displaying the same 25 bases deletion in the D genome allele of GBSS. Two tetraploids and five hexaploids were null 4A for GBSS. There was little difference in the amylose contents and amylose-lipid dissociation peak temperatures of the synthetic hexaploids and the lower ploidy wheats. The synthetic hexaploids showed intermediate swelling power values with the durums giving the highest swelling powers. The durums also had higher B granule contents than the A. tauschii accessions, but not as high as the synthetics. However, the A. tauschii samples gave the highest gelatinisation peak temperatures. The presence of the null 4A mutation was positively correlated with swelling power, amylose content and DSC measurements. The new smaller D genome allele of GBSS was associated with slightly higher swelling power. These results confirm the value of wheat progenitor lines as sources of new starch properties for hexaploid wheat. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.